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    11 views78 pages

    Histolopathology 13078 (11819)

    Uploaded by

    Rishikesh Doke

    Copyright:

    © All Rights Reserved
    Download as PDF or read online from Scribd
    Flag for inappropriate content
    of 78
    = _ Biopsy sample collectionmethods. —-+ © A biopsy is a medical procedure where a sample of tissue is removed from the body for further examination under a microscope. There are several methods of collecting biopsy samples, depending on the type of tissue being sampled and the location of the tissue in the body. Here are some common methods of biopsy sample collection: 1. Needle biopsy: A needle is inserted through the skin into the tissue being sampled, and a small sample of tissue is extracted using suction or cutting. Needle biopsies are commonly used for tumors or’ ee = Biopsy sample collection methods. -+ cysts that can be felt through the skin, or for certain organs such as the liver or kidney. 2. Incisional biopsy: A small incision is made in the skin, and a sample of tissue is removed using surgical instruments. Incisional biopsies are commonly used for larger tumors or for tissues that cannot be reached by needle biopsy. 3. Excisional biopsy: The entire abnormal tissue is removed, along with a margin of normal tissue around it. Excisional biopsies are commonly used for small tumors or for superficial skin lesions. 4. Endoscopic biopsy: An endoscope, which ic athin flevihle tiihe with a = Biopsysamplecollectionmethods. + camera and a light on the end, is inserted through a natural opening in the body or a small incision in the skin. The endoscope is used to visualize the tissue being sampled, and asmall sample is removed using forceps or a biopsy needle. Endoscopic biopsies are commonly used for tissues inside the gastrointestinal tract, the respiratory system, or the urinary Vitctaan 5. Bone marrow biopsy: A special needle is inserted into the bone marrow, usually at the hip, anda small sample of bone marrow is removed. Bone marrow biopsies = _ Biopsy sample collectionmethods. + is removed using a punch biopsy tool or a scalpel. Skin biopsies are commonly used to diagnose skin conditions such as melanoma or other skin cancers. The choice of biopsy method depends on several factors, including the location of the tissue being sampled, the size and type of the lesion, and the patient's medical history and overall health. The specific biopsy method chosen by a doctor will depend on these factors and the specific needs of the patient. Oo fb & CryoSat & Frozen Sections =F coy CryoSat is a European Space Agency (ESA) satellite mission that was launched in 2010 to measure changes in the thickness of the Earth's polar ice sheets and sea ice. The satellite uses a sophisticated radar altimeter to measure the height of the ice surface with a high degree of accuracy. By comparing these measurements over time, scientists can determine how the ice sheets and sea ice are changing and how they are contributing to sea level rise. Frozen sections are thin slices of tissue that are cut from a frozen sample using a special instrument CryoSat & Frozen Sections called a cryostat. The tissue is typically frozen to preserve its structure and prevent degradation of its cellular components. Frozen sections are commonly used in medical research and pathology to examine tissues under a microscope and diagnose diseases. The process of cutting frozen sections involves mounting the frozen tissue sample onto a cutting surface and slicing it into thin sections using a microtome. The resulting sections can be stained and examined under a microscope to reveal the structure and composition of the tissue. NY Od & = Mounting a Stained Section. + coy The procedure for mounting a section after staining can vary depending on the specific staining technique used and the purpose of the staining. However, a general procedure for mounting a section after staining is as follows: 1. Remove the slide with the stained section from the staining solution and rinse it in distilled water to igTa nleY{< Hela => Cees Kola 2. Dip the slide in a container of alcohol (usually 70% ethanol or isopropanol) to dehydrate the LASSI 3. Clear the section by transferring iv to acontainer of clearina aaent Mounting a Stained Section. a such as xylene or toluene. This step removes any remaining alcohol and makes the tissue more transparent. . Transfer the section to a slide with a drop of mounting medium. A commonly used mounting medium is Canada balsam, but other types of mounting media can be used as well. . Gently place a coverslip over the section using a pair of forceps, making sure that there are no air bubbles trapped between the coverslip and the section. . Press down lightly on the coverslip with a flat object, such as the ~ Mounting a Stained Section. a handle of a scalpel or a glass rod, to distribute the mounting medium evenly and remove any excess. 7. Seal the edges of the coverslip with a clear nail polish or a commercial mounting medium to prevent the coverslip from moving or drying out. 8. Label the slide with the stain used and any other relevant information. 9. Allow the mounting medium to dry completely before examining the slide under a microscope. It is important to note that the specific steps and techniques used may vary depending on the staining Stel only and the type of tissue being Hl Mounting a Stained Section. so or drying out. 8. Label the slide with the stain used and any other relevant information. 9. Allow the mounting medium to dry completely before examining the slide under a microscope. It is important to note that the specific steps and techniques used may vary depending on the staining procedure and the type of tissue being examined. It is also important to follow any specific protocols or guidelines provided by the staining procedure or the laboratory. ie = Hormonal evaluation methods. a © Staining methods are not typically used for hormonal evaluation as hormones are typically measured using immunoassays or other laboratory techniques that do not require staining. However, there are some situations in which staining may be used to evaluate hormones, such as in the evaluation of hormone- producing tumors. One example of a staining method that may be used to evaluate hormones is immunohistochemistry (IHC). IHC involves the use of antibodies that are specific to the hormone of interest. These antibodies aro lahaload with a cArnamananir nr Hormonal evaluation methods. a are labeled with a chromogenic or fluorescent tag that allows them to be visualized under a microscope. Tissue samples are collected and prepared on slides, then treated with the labeled antibodies. The antibodies bind to the hormone of interest in the tissue sample, and the resulting staining pattern can be visualized and evaluated. Another staining method that may be used to evaluate hormones is radioimmunoassay (RIA). RIA involves the use of a radiolabeled hormone and an antibody that is specific to the hormone of interest. The radiolabeleu harmanna ramnataciwith tha harmana Hormonal evaluation methods. a hormone of interest. The radiolabeled hormone competes with the hormone in the sample for binding to the antibody. The amount of radiolabeled hormone that is bound to the antibody is measured, and this measurement is used to calculate the amount of hormone in the sample. Overall, staining methods are not typically used for hormonal evaluation, but in certain situations, they may be employed to visualize and evaluate hormones in tissue samples. i =~ Gynaecological Sample Collection. SF 6 Gynecological samples are usually collected during a routine gynecological examination or for diagnostic purposes. The most common types of gynecological samples collected are: 1. Pap Smear: This is a screening test for cervical cancer. The healthcare provider uses a speculum to visualize the cervix and collect cells from the surface of the cervix using asmall brush or spatula. 2. Vaginal Swab: A sterile cotton swab is used to collect a sample of discharge from the vagina. This NY sample is then tested for infections Gynaecological Sample Collection. such as yeast, bacterial vaginosis, or sexually transmitted infections. 3. Endometrial Biopsy: A sample of the lining of the uterus is taken using a small instrument called a biopsy curette. This procedure is usually done to investigate abnormal uterine bleeding or to diagnose endometrial cancer. 4. Colposcopy: A colposcopy is a diagnostic procedure used to examine the cervix, vagina, and vulva. A speculum is used to hold Se open the vagina, and a colposcope is used to visualize the area. A biopsy may be taken during this procedure if abnormal cells are = Gynaecological Sample Collection. + biopsy may be taken during this procedure if abnormal cells are identified. 5. Cervical Culture: A swab is taken from the cervix to test for bacterial or viral infections such as chlamydia, gonorrhea, or human papillomavirus (HPV). It is important to follow your healthcare provider's instructions carefully when preparing for these procedures, and to discuss any concerns or questions you may have with them beforehand. a = Staining types explained. © Staining is acommon laboratory technique used to enhance the contrast of biological specimens, making them easier to visualize and study. There are several types of staining techniques, which are used depending on the type of specimen and the information that needs to be obtained. 1. Simple Staining: This is the most basic staining technique and involves the use of a single stain such as crystal violet or methylene blue, which binds to all parts of the specimen equally, giving it a ’ Vv uniform color. Staining types explained. + 2. Differential Staining: This technique involves the use of two or more stains, each of which interacts differently with different parts of the specimen, highlighting specific features. The most common type of differential staining is the Gram stain, which is used to differentiate between Gram-positive and Gram-negative bacteria. . Acid-Fast Staining: This technique is used to identify acid-fast bacteria such as Mycobacterium tuberculosis, which are resistant to traditional staining methods. The bacteria are first stained with Staining types explained. = carbol fuchsin, and then decolorized with acid-alcohol, before being counterstained with methylene blue. . Immunohistochemistry: This technique uses antibodies to detect specific proteins or other molecules in tissues. The antibodies are labeled with a fluorescent or enzyme tag, which can be visualized under a microscope. . Fluorescent Staining: This technique uses fluorescent dyes to label specific structures in cells, tissues or organisms. The dyes absorb light at one wavelength Pine Pe Sa ae ey < Se | Perry ape ey | Snee Overall, the choice of staining Staining types explained. ee ee gyre ee emit it at another, allowing for the visualization of structures in living cells. . Histochemical Staining: This technique is used to visualize specific biochemical compounds in tissues. The staining involves the use of specific dyes that react with the compound of interest, producing a visible color change. Examples of histochemical staining include the periodic acid- Schiff (PAS) stain, which is used to detect glycogen and other carbohydrates in tissues. ao Staining types explained. in tissues. The staining involves the use of specific dyes that react with the compound of interest, producing a visible color change. Examples of histochemical staining include the periodic acid- Schiff (PAS) stain, which is used to detect glycogen and other carbohydrates in tissues. Overall, the choice of staining technique depends on the nature of the specimen, the question being asked, and the equipment and resources available. Histology Importance 6 Histology is the study of the microscopic structure of tissues and organs, while histopathology is the study of abnormal changes in tissues and organs that are caused by disease. In histology, tissues are typically examined under a microscope after being processed and stained to reveal their structure, while in histopathology, diseased tissues are examined to determine the underlying cause of the disease. The importance of histology and histopathology lies in their ability to provide information about the structure and function of tissues and Histology Importance + organs, as well as their ability to diagnose diseases. Histology is used in a wide range of fields, including medicine, veterinary medicine, biology, and forensic science, to name a few. It can help identify different types of cells, their arrangement, and the structure of organs and tissues. Histopathology, on the other hand, is crucial in the diagnosis and treatment of diseases, as it can reveal changes in tissues that are indicative of diseases such as cancer, infections, and autoimmune disorders. It can also provide important information about the stage and severity of a disease, which is essential for selecting Histology Importance oP in tissues that are indicative of diseases such as cancer, infections, and autoimmune disorders. It can also provide important information about the stage and severity of a disease, which is essential for selecting appropriate treatment options. In summary, histology and histopathology are important fields of study that help us understand the structure and function of tissues and organs, diagnose diseases, and develop effective treatment strategies. = Fixatives: Ideal Criteria a © Fixatives are chemical solutions that are used to preserve biological specimens by preventing the degradation of tissues and cells. The primary function of fixatives is to crosslink and stabilize proteins and other cellular components, preventing autolysis and putrefaction. The ideal fixatives should meet the following criteria: 1. Preserve cellular morphology: The fixative should maintain the natural structure and shape of the tissue or cell, without causing See distortion or shrinkage. Fixatives: Ideal Criteria Se . Cross-linkation: The fixative should create stable chemical bonds between the cellular components to prevent degradation. . Rapid penetration: The fixative should be able to penetrate the tissue or cell quickly and uniformly, ensuring even distribution and preservation of all components. . Compatibility with downstream processing: The fixative should not interfere with subsequent staining, imaging, or molecular analyses of the specimen. . Long-term preservation: The fixative should maintain the stability of the specimen for an a avtandnd narinA anshlina lana Cla De eS 2 ae F ite nr ae Be a tars F Fixatives: Ideal Criteria fixative should maintain the stability of the specimen for an extended period, enabling long- term storage and analysis. Commonly used fixatives in biology include formaldehyde, glutaraldehyde, paraformaldehyde, ethanol, methanol, and acetone, each with their own advantages and disadvantages. The choice of fixative depends on the nature of the sample, the intended downstream applications, and the preservation needs. a Vacuum Impregnation Oven Working -+ coy A vacuum impregnation oven is a specialized piece of equipment used in the process of impregnating porous Are Ll gre lCMSLU Lol pe ToMexole-Tanl (xm aa Ne=) liquid or resin. The basic principle behind this process is to remove the air and any other volatile components from the porous material, thereby creating a void or space within it. This void can then be filled with a liquid or resin, which will harden or solidify, creating a more durable and resilient material. The vacuum impregnation oven typically consists of a chamber or NeTSI—1 Maal leva mal oMaarelXclat-l Ke el-1 = Vacuum |Impregnation Oven Working + impregnated is placed. The chamber is then sealed and a vacuum is created, using a vacuum pump or other similar equipment. This vacuum removes the air and any other volatile components from the material, (ol g-F-ldl ate m-MYeO)(o mM TIMA Lane Once the vacuum has been established, the liquid or resin to be used for impregnation is introduced into the chamber, either by flooding or by slowly filling the chamber to a predetermined level. The vacuum draws the liquid or resin into the voids within the material, ensuring that it completely fills any empty spaces ard rraatac aiinifarm and erancictant = Vacuum Impregnation Oven Working + creates a uniform and consistent coating. After the impregnation process is complete, the chamber is slowly re- pressurized, allowing the liquid or resin to solidify or harden within the material. The impregnated material is then removed from the chamber, cleaned and dried, and is ready for use. In summary, the vacuum impregnation oven is a highly specialized piece of equipment that uses vacuum pressure to remove air and volatile components from elt materials, allowing them to be = Vacuum Impregnation Oven Working + resin to solidify or harden within the material. The impregnated material is then removed from the chamber, cleaned and dried, and is ready for use. In summary, the vacuum impregnation oven is a highly specialized piece of equipment that uses Vacuum pressure to remove air and volatile components from porous materials, allowing them to be impregnated with liquid or resin, creating a stronger and more durable material. od & Pap Stain Procedure + © The Papanicolaou stain, also known as Pap stain, is a multichromatic staining technique used to visualize cells and tissue structures under a microscope. The principle of the Pap stain is to differentiate various cellular components based on their chemical properties and affinity to different Selale The Papanicolaou stain consists of three main reagents: hematoxylin, eosin, and a mixture of orange G and light green. Hematoxylin stains acidic structures such as nuclei blue-purple while eosin stains basic structures like cytoplasm pink-red. The orange G Pap Stain Procedure and light green mixture helps to highlight the intermediate filaments of cells. The staining procedure of the Pap stain usually involves the following Sic] 0)

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